PDF(Pubmed)
Abstract：
The widely used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas) system is thought to have evolved from IS200/IS605 transposons. TnpB proteins, encoded by one type of IS200/IS605 transposon, are considered to be the evolutionary ancestors of Cas12 nucleases, which have been engineered to function as RNA-guided DNA endonucleases for genome editing in bacteria and human cells. TnpB nucleases, which are smaller than Cas nucleases, have been engineered for use in genome editing in animal systems, but the feasibility of this approach in plants remained unknown. Here, we obtained stably transformed genome-edited mutants in rice (Oryza sativa) by adapting three recently identified TnpB genome editing vectors, encoding distinct TnpB nucleases (ISAam1, ISDra2, and ISYmu1), for use in plants, demonstrating that the hypercompact TnpB proteins can effectively edit plant genomes. ISDra2 and ISYmu1 precisely edited their target sequences, with no off-target mutations detected, showing that TnpB transposon nucleases are suitable for development into a new genome editing tool for plants. Future modifications improving the genome-editing efficiency of the TnpB system will facilitate plant functional studies and breeding programs.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s42994-024-00172-6.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s42994-024-00172-6.
摘要：
广泛使用的成簇的规则间隔短回文重复序列（CRISPR）/CRISPR相关核酸酶（Cas）系统被认为是从IS200/IS605转座子进化而来的。TnpB蛋白,由一种类型的IS200/IS605转座子编码，被认为是Cas12核酸酶的进化祖先，这些酶被设计为RNA指导的DNA核酸内切酶，用于细菌和人类细胞的基因组编辑。TnpB核酸酶，比Cas核酸酶小，已经被设计用于动物系统的基因组编辑，但是这种方法在植物中的可行性仍然未知。这里,通过采用三个最近鉴定的TnpB基因组编辑载体，我们获得了水稻（Oryzasativa）中稳定转化的基因组编辑突变体，编码不同的TnpB核酸酶（ISAam1，ISDra2和ISYmu1），用于植物，证明超紧密TnpB蛋白可以有效编辑植物基因组。ISDra2和ISYmu1精确编辑了它们的靶序列，没有检测到脱靶突变，表明TnpB转座子核酸酶适合开发为植物的新基因组编辑工具。改善TnpB系统基因组编辑效率的未来修改将促进植物功能研究和育种计划。
■在线版本包含补充材料，可在10.1007/s42994-024-00172-6获得。
■在线版本包含补充材料，可在10.1007/s42994-024-00172-6获得。
